Zinc finger and BTB domain-containing protein 46 is essential for survival and proliferation of acute myeloid leukemia cell line but dispensable for normal hematopoiesis
Zinc finger and BTB domain-containing protein 46 is essential for survival and proliferation of acute myeloid leukemia cell line but dispensable for normal hematopoiesis
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含有锌指和 BTB 结构域的蛋白 46 对于急性髓性白血病细胞系的生存和增殖至关重要，但对于正常造血来说不是必需的
DOI：
10.1097/cm9.0000000000000878
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发表时间：
2020-07-20
影响因子：
6.1
通讯作者：
Hou, Yu
中科院分区：
文献类型：
--
作者：
Liu, Yuan-Yuan;Xiao, Fei-Fei;Hou, Yu
背景 含锌指和 BTB 结构域的蛋白 46 (Zbtb46) 是在经典树突状细胞中鉴定的转录因子，并维持树突状细胞静止在稳定状态。Zbtb46 据报道是急性髓系白血病 (AML) 的阴性指标。我们发现，与成熟细胞相比，Zbtb46 在造血干细胞和祖细胞 (HSPC) 中的表达水平相对较高，与正常骨髓 (BM) 细胞相比，在 AML 细胞中的表达水平较高。然而，Zbtb46 在 HSPC 和 AML 细胞中的作用仍不清楚。因此，我们试图阐明Zbtb46在正常造血和AML细胞中的作用。方法我们生成了Zbtb46(fl/fl)和Zbtb46(fl/fl)Mx1-Cremice。通过腹腔注射双链聚(I)诱导Zbtb46(fl/fl)Mx1-Cremice中Zbtb46的缺失。聚(C) (poly(I:C))，简称Zbtb46cKO。确认Zbtb46缺失后，通过流式细胞仪分析HSPC和成熟血细胞的频率和数量。连续腹腔注射5-氟尿嘧啶测定Zbtb46(fl/fl)和Zbtb46cKO小鼠HSC的增殖能力。使用癌症基因组图谱的数据分析了 Zbtb46 表达与预后之间的相关性。为了研究Zbtb46在AML细胞中的作用，我们使用表达针对Zbtb46的小发夹RNA的慢病毒载体敲低了THP-1细胞中Zbtb46的表达。通过细胞计数测定测定细胞增殖率。通过流式细胞术测定细胞凋亡和溴脱氧尿苷掺入。结果 Zbtb46cKO 小鼠及其 Zbtb46(fl/fl) 同窝小鼠中 HSPC 和成熟血细胞的百分比和绝对数量相当（Zbtb46(fl/fl) 与 Zbtb46cKO，HPC：801,310 +/- 84,282vs.907,202 +/- 97,403，t= 0.82，P=0.46； LSK：86,895 +/- 7802vs.102,210 +/- 5025，t = 1.65，P = 0.17； HSC：19,753 +/- 3116 对比 17,608 +/- 3508，t = 0.46，P = 0.67）。来自Zbtb46(fl/fl)Mx1-Cremice的HSC的再增殖能力与来自Zbtb46(fl/fl)对照的HSC相似(P = 0.26)。与来自正常对照的总BM细胞相比，Zbtb46在AML细胞中的表达升高。 THP-1细胞中Zbtb46的敲低导致细胞凋亡显着增加并减少细胞生长和增殖。结论 总的来说，我们的数据表明 Zbtb46 对于 AML 细胞的存活和增殖至关重要，但对于正常造血来说是可有可无的。
Background Zinc finger and BTB domain-containing protein 46 (Zbtb46) is a transcription factor identified in classical dendritic cells, and maintains dendritic cell quiescence in a steady state.Zbtb46has been reported to be a negative indicator of acute myeloid leukemia (AML). We found thatZbtb46was expressed at a relatively higher level in hematopoietic stem and progenitor cells (HSPCs) compared to mature cells, and higher in AML cells compared to normal bone marrow (BM) cells. However, the role ofZbtb46in HSPCs and AML cells remains unclear. Therefore, we sought to elucidate the effect ofZbtb46in normal hematopoiesis and AML cells. Methods We generatedZbtb46(fl/fl)andZbtb46(fl/fl)Mx1-Cremice. The deletion ofZbtb46inZbtb46(fl/fl)Mx1-Cremice was induced by intraperitoneal injection of double-stranded poly (I). poly (C) (poly(I:C)), and referred asZbtb46cKO. After confirming the deletion ofZbtb46, the frequency and numbers of HSPCs and mature blood cells were analyzed by flow cytometry. Serial intraperitoneal injection of 5-fluorouracil was administrated to determine the repopulation ability of HSCs fromZbtb46(fl/fl)andZbtb46cKO mice. The correlation betweenZbtb46expression and prognosis was analyzed using the data from the Cancer Genome Atlas. To investigate the role ofZbtb46in AML cells, we knocked down the expression ofZbtb46in THP-1 cells using lentiviral vectors expressing small hairpin RNAs targetingZbtb46. Cell proliferation rate was determined by cell count assay. Cell apoptosis and bromodeoxyuridine incorporation were determined by flow cytometry. Results The percentages and absolute numbers of HSPCs and mature blood cells were comparable inZbtb46cKO mice and itsZbtb46(fl/fl)littermates (Zbtb46(fl/fl)vs. Zbtb46cKO, HPC: 801,310 +/- 84,282vs.907,202 +/- 97,403,t= 0.82,P= 0.46; LSK: 86,895 +/- 7802vs.102,210 +/- 5025,t = 1.65,P = 0.17; HSC: 19,753 +/- 3116vs.17,608 +/- 3508,t = 0.46,P = 0.67). The repopulation ability of HSCs fromZbtb46(fl/fl)Mx1-Cremice was similar to those fromZbtb46(fl/fl)control (P = 0.26).Zbtb46had elevated expression in AML cells compared to total BM cells from normal control. Knockdown ofZbtb46in THP-1 cells led to a significant increase in cell apoptosis and reduced cell growth and proliferation. Conclusion Collectively, our data indicate thatZbtb46is essential for survival and proliferation of AML cells, but dispensable for normal hematopoiesis.